Acute myocardial infarction (MI) and coronary artery disease (CAD) continue to be the leading cause of death and disability in the world despite recent diagnostic and treatment advances. Moreover, this trend is expected to dramatically worsen given global life expectancy increases, population growth, and the rapidly escalating prevalence of obesity and diabetes. Current methods of detection for CAD is easily diagnosed through routine stress testing and coronary angiography, however catastrophes such as MI remain highly unpredictable despite well known risk factors. Circulating endothelial cells (CECs) and endothelial derived microparticles (CMPs) have considerable promise as a biomarker for acute Myocardial Infarction. It has demonstrated that CECs were found in the blood at significant levels in individuals at high risk for or those who already had an acute myocardial infarction, but not in individuals with stable CAD. It has also been reported that the elevated levels of CFC-DNA is a strong prognostic marker in acute MI. Current methods for isolated CFC-DNA and CMPs are cumbersome, destructive and unreliable. The amount of cell-free DNA in healthy patients varies from 12.5 to 60.1 ng/mL and in MI patients from 113 to 909 ng/mL. Markers and antibody stains for identifying EMPs include CD144, von Willebrand Factor (vWF) and Ulex Europaeus Agluttin (UEA), a lectin stain. Our prototype device will enable POC immediate prognostic information at a reasonable cost once the FDA approves it. This project will focus on the pre-clinical aspect of using a proprietary prototype ACE device to isolate EMPs and CFC-DNA from MI patient blood and analyzing to reveal further knowledge and understanding. The goal is to now develop a robust point-of-care MI diagnostic system, which will separate CFC-HMW DNA and EMPs directly from whole blood and then elute it for further analysis using gel electrophoresis, qRT-PCR and fluorescence analysis The specific aims for this Phase I application are: 1 - Use prototype ACE device to characterize performance for fluorescent beads and lysed blood. 2 - Use prototype ACE device to characterize performance for MI plasma and compare against gold standards (QIAamp kit for CFC-DNA, FACS for EMPs). 3 - Use prototype ACE device to isolate EMPs and CFC-DNA from 10 controls and 10 MI patients and compare to CEC data identified for the same patients by Scripps Health. The SBIR Phase I grant will allow the construction of this device, which will further enable rapid translation for manufacturing and commercialization. A future Phase II proposal will modify the prototype as needed, and will construct an Analytical System, correlate fresh whole blood vs. previously frozen plasma, and integrate PCR amplification for identification of endothelial markers through fluorescence and genotyping PUBLIC HEALTH RELEVANCE: This project is directed at the development of a point-of-care heart attack diagnostic system that will allow rapid, cost effective screening for all Americans. The system does this by rapidly isolating cell free circulating DNA and endothelial microparticles in blood, an early heart attack biomarker that has a growing consensus, using a prototype AC Electrokinetic device. Further development of this device and biomarker will create a new point-of-care heart attack diagnostic device that will lead to better quality healthcare for all Americans as well as lowering costs associated with heart attacks through early detection and subsequent treatment.